Stabilized Formulations Containing Anti-Interleukin-6 Receptor (IL-6R) Antibodies

ABSTRACT

The present invention provides pharmaceutical formulations comprising a human antibody that specifically binds to human interleukin-6 receptor (hIL-6R). The formulations may contain, in addition to an anti-hIL-6R antibody, at least one amino acid, at least one sugar, and/or at least one non-ionic surfactant. The pharmaceutical formulations of the present invention exhibit a substantial degree of antibody stability after storage for several months.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application is a continuation of U.S. application Ser. No.12/986,223, filed Jan. 7, 2011, which claims the benefit under 35 USC119(e) of U.S. Application No. 61/293,227, filed Jan. 8, 2010, each ofwhich is incorporated herein by reference in its entirety.

REFERENCE TO A SEQUENCE LISTING

This application includes an electronic sequence listing in a file named“465070-Sequence.txt”, created on Sep. 22, 2015 and containing 37,428bytes, which is hereby incorporated by reference in its entirety for allpurposes.

FIELD OF THE INVENTION

The present invention relates to the field of therapeutic antibodyformulations. More specifically, the present invention relates to thefield of pharmaceutical formulations comprising a human antibody thatspecifically binds to human interleukin-6 receptor.

BACKGROUND

Therapeutic macromolecules (e.g., antibodies) must be formulated in amanner that not only makes the molecules suitable for administration topatients, but also maintains their stability during storage. Forexample, therapeutic antibodies in liquid solution are prone todegradation, aggregation and/or undesired chemical modifications unlessthe solution is formulated properly. The stability of an antibody inliquid formulation depends not only on the kinds of excipients used inthe formulation, but also on the amounts and proportions of theexcipients relative to one another. Furthermore, other considerationsaside from stability must be taken into account when preparing a liquidantibody formulation. Examples of such additional considerations includethe viscosity of the solution and the concentration of antibody that canbe accommodated by a given formulation. Thus, when formulating atherapeutic antibody, great care must be taken to arrive at aformulation that remains stable, contains an adequate concentration ofantibody, and possesses a suitable viscosity as well as other propertieswhich enable the formulation to be conveniently administered topatients.

Antibodies to the human interleukin-6 receptor (hIL-6R) are one exampleof a therapeutically relevant macromolecule that requires properformulation. Anti-hIL-6R antibodies are clinically useful for thetreatment and/or prevention of diseases such as rheumatoid arthritis,ankylosing spondylitis, and other conditions. Exemplary anti-IL-6Rantibodies are described, inter alia, in U.S. Pat. Nos. 7,582,298;6,410,691; 5,817,790; 5,795,695; and 6,670,373. A particularly importantanti-hIL-6R antibody with great therapeutic potential is the antibodyreferred to in U.S. Pat. No. 7,582,298 as VQ8F11-21 (also referred toherein as “mAb1”).

Although anti-hIL-6R antibodies are known, there remains a need in theart for novel pharmaceutical formulations comprising anti-hIL-6Rantibodies which are sufficiently stable and also suitable foradministration to patients.

BRIEF SUMMARY OF THE INVENTION

The present invention satisfies the aforementioned need by providingpharmaceutical formulations comprising a human antibody thatspecifically binds to human interleukin-6 receptor (hIL-6R). Theformulations of the invention may comprise excipients in addition to theanti-hIL-6R antibody. For example, in certain embodiments, theformulation may comprise (i) a human antibody that specifically binds tohIL-6R; (ii) at least one amino acid; and (iii) at least onecarbohydrate. The amino acid can be, e.g., histidine and/or arginine.The carbohydrate can be a sugar such as, e.g., sucrose, glucose,mannitol, lactose or trehalose.

According to certain embodiments of the present invention, theformulation further comprises a non-ionic surfactant. The non-ionicsurfactant can be, e.g., polysorbate 20, polysorbate 80, polyoxyethylenesorbitan monooleate, polyethylene glycol, etc.

The antibody contained within the pharmaceutical formulations of thepresent invention can be any antibody which specifically binds tohIL-6R. Exemplary antibodies that may be contained within theformulations of the invention are antibodies comprising a heavy chainvariable region (HCVR) and a light chain variable region (LCVR), whereinthe HCVR comprises a heavy chain complementary determining region (HCDR)1 having the amino acid sequence of SEQ ID NO: 20, a HCDR2 having theamino acid sequence of SEQ ID NO:22, and a HCDR3 having the amino acidsequence of SEQ ID NO:24; and wherein the LCVR comprises a light chaincomplementary determining region (LCDR) 1 having the amino acid sequenceof SEQ ID NO: 28, a LCDR2 having the amino acid sequence of SEQ IDNO:30, and a LCDR3 having the amino acid sequence of SEQ ID NO:32. Incertain embodiments, the antibody contained within the formulations ofthe present invention are antibodies comprising a HCVR having the aminoacid sequence of SEQ ID NO:18 and a LCVR having the amino acid sequenceof SEQ ID NO:26.

The antibody formulations of the present invention may be containedwithin any suitable container useful for storing pharmaceuticalformulations. Examples of such suitable containers include, e.g., glassor plastic vials, syringes and cartridges. The container may be clear oropaque (e.g., amber colored).

According to certain aspects of the present invention, thepharmaceutical formulations remain relatively stable following storagefor several days, months or years at a given temperature. For example,in certain exemplary embodiments of the present invention, a highpercentage of the antibody (e.g., 90%, 95%, 96% or more) is maintainedin its native form following at least 3, 6, 9 or more months of storage.The percentage of native form of the antibody may be measured, e.g., bySE-HPLC, or by any other method known in the art. The storagetemperature at which stability of the antibody is maintained can be,e.g., −80° C., −40° C., −20° C., 0° C., 5° C., 25° C., 45° C., orhigher.

Other embodiments of the present invention will become apparent from areview of the ensuing detailed description.

BRIEF DESCRIPTION OF THE FIGURES

FIG. 1 shows the percent of native mAb1 remaining, as measured bySE-HPLC, following various amounts of time of storage at −20° C. (filledtriangles), −30° C. (filled squares), and −80° C. (filled diamonds).

FIG. 2 shows the percent of acidic species of mAb1, as measured byCEX-HPLC, following various amounts of time of storage at −20° C.(filled triangles), −30° C. (filled squares), and −80° C. (filleddiamonds).

FIG. 3 shows the percent of native mAb1 remaining in various minimalexcipient formulations, as measured by SE-HPLC, following variousamounts of time of storage at −30° C. Filled diamonds representformulation 1 (80 mg/mL mAb1, 0.13% polysorbate 20, 6% sucrose, 10 mMhistidine); filled squares represent formulation 2 (80 mg/mL mAb1, 0.13%polysorbate 20, 10 mM histidine); filled triangles represent formulation3 (80 mg/mL mAb, 1% sucrose, 10 mM histidine); open squares representformulation 4 (80 mg/mL mAb1, 2% sucrose, 10 mM histidine); asterisksrepresent formulation 5 (80 mg/mL mAb1, 4% sucrose, 10 mM histidine);filled circles represent formulation 6 (80 mg/mL mAb1, 6% sucrose, 10 mMhistidine); crosses represent formulation 7 (80 mg/mL antibody, 10 mMhistidine); and open circles represent formulation 8 (65 mg/mL antibody,10 mM histidine). All formulations are set out in Table 6 (see Example2, below).

FIG. 4 shows the percent of native mAb1 remaining in various minimalexcipient formulations, as measured by SE-HPLC, following variousamounts of time of storage at −20° C. Filled diamonds representformulation 1 (80 mg/mL mAb1, 0.13% polysorbate 20, 6% sucrose, 10 mMhistidine); filled squares represent formulation 2 (80 mg/mL mAb1, 0.13%polysorbate 20, 10 mM histidine); filled triangles represent formulation3 (80 mg/mL mAb, 1% sucrose, 10 mM histidine); open squares representformulation 4 (80 mg/mL mAb1, 2% sucrose, 10 mM histidine); asterisksrepresent formulation 5 (80 mg/mL mAb1, 4% sucrose, 10 mM histidine);filled circles represent formulation 6 (80 mg/mL mAb1, 6% sucrose, 10 mMhistidine); crosses represent formulation 7 (80 mg/mL antibody, 10 mMhistidine); and open circles represent formulation 8 (65 mg/mL antibody,10 mM histidine). All formulations are set out in Table 6 (see Example2, below).

DETAILED DESCRIPTION

Before the present invention is described, it is to be understood thatthis invention is not limited to particular methods and experimentalconditions described, as such methods and conditions may vary. It isalso to be understood that the terminology used herein is for thepurpose of describing particular embodiments only, and is not intendedto be limiting, since the scope of the present invention will be limitedonly by the appended claims.

Unless defined otherwise, all technical and scientific terms used hereinhave the same meaning as commonly understood by one of ordinary skill inthe art to which this invention belongs. As used herein, the term“about,” when used in reference to a particular recited numerical value,means that the value may vary from the recited value by no more than 1%.For example, as used herein, the expression “about 100” includes 99 and101 and all values in between (e.g., 99.1, 99.2, 99.3, 99.4, etc.).

Although any methods and materials similar or equivalent to thosedescribed herein can be used in the practice or testing of the presentinvention, the preferred methods and materials are now described. Allpublications mentioned herein are incorporated herein by reference todescribe in their entirety.

PHARMACEUTICAL FORMULATIONS

As used herein, the expression “pharmaceutical formulation” means acombination of at least one active ingredient (e.g., a small molecule,macromolecule, compound, etc. which is capable of exerting a biologicaleffect in a human or non-human animal), and at least one inactiveingredient which, when combined with the active ingredient and/or one ormore additional inactive ingredients, is suitable for therapeuticadministration to a human or non-human animal. The term “formulation,”as used herein, means “pharmaceutical formulation” unless specificallyindicated otherwise. The present invention provides pharmaceuticalformulations comprising at least one therapeutic polypeptide. Accordingto certain embodiments of the present invention, the therapeuticpolypeptide is an antibody that binds specifically to humaninterleukin-6 receptor (hIL-6R) or an antigen-binding fragment thereof.More specifically, the present invention includes pharmaceuticalformulations that comprise: (i) a human antibody that specifically bindsto hIL-6R; (ii) histidine; and (iii) a carbohydrate. Additionalcomponents may be included in the formulations of the present inventionsuch as, e.g., at least one non-ionic surfactant, and at least oneadditional amino acid. Specific exemplary components and formulationsincluded within the present invention are described in detail below.

The pharmaceutical formulations of the present invention may, in certainembodiments, be fluid formulations. As used herein, the expression“fluid formulation” means a mixture of at least two components thatexists predominantly in the fluid state at about 5° C. to about 45° C.Fluid formulations include, inter alia, liquid formulations. Fluidformulations may be of low, moderate or high viscosity depending ontheir particular constituents.

ANTIBODIES THAT BIND SPECIFICALLY TO hIL-6R

The pharmaceutical formulations of the present invention may comprise ahuman antibody, or an antigen-binding fragment thereof, that bindsspecifically to hIL-6R. As used herein, the term “hIL-6R” means a humancytokine receptor that specifically binds interleukin-6 (IL-6). Incertain embodiments, the antibody contained within the pharmaceuticalformulations of the present invention binds specifically to theextracellular domain of hIL-6R. The extracellular domain of hIL-6R isrepresented by the amino acid sequence of SEQ ID NO:74.

The term “antibody”, as used herein, is generally intended to refer toimmunoglobulin molecules comprising four polypeptide chains, two heavy(H) chains and two light (L) chains inter-connected by disulfide bonds,as well as multimers thereof (e.g., IgM); however, immunoglobulinmolecules consisting of only heavy chains (i.e., lacking light chains)are also encompassed within the definition of the term “antibody.” Eachheavy chain comprises a heavy chain variable region (abbreviated hereinas HCVR or VH) and a heavy chain constant region. The heavy chainconstant region comprises three domains, CH1, CH2 and CH3. Each lightchain comprises a light chain variable region (abbreviated herein asLCVR or VL) and a light chain constant region. The light chain constantregion comprises one domain (CL1). The VH and VL regions can be furthersubdivided into regions of hypervariability, termed complementarydetermining regions (CDRs), interspersed with regions that are moreconserved, termed framework regions (FR). Each VH and VL is composed ofthree CDRs and four FRs, arranged from amino-terminus tocarboxy-terminus in the following order: FR1, CDR1, FR2, CDR2, FR3,CDR3, FR4.

Unless specifically indicated otherwise, the term “antibody,” as usedherein, shall be understood to encompass complete antibody molecules aswell as antigen-binding fragments thereof. The term “antigen-bindingportion” or “antigen-binding fragment” of an antibody (or simply“antibody portion” or “antibody fragment”), as used herein, refers toone or more fragments of an antibody that retain the ability tospecifically bind to hIL-6R.

An “isolated antibody”, as used herein, is intended to refer to anantibody that is substantially free of other antibodies having differentantigenic specificities (e.g., an isolated antibody that specificallybinds hIL-6R is substantially free of antibodies that specifically bindantigens other than hIL-6R).

The term “specifically binds,” or the like, means that an antibody orantigen-binding fragment thereof forms a complex with an antigen that isrelatively stable under physiologic conditions. Specific binding can becharacterized by a dissociation constant of at least about 1×10⁻⁶ M orgreater. Methods for determining whether two molecules specifically bindare well known in the art and include, for example, equilibriumdialysis, surface plasmon resonance, and the like. An isolated antibodythat specifically binds hIL-6R may, however, have cross-reactivity toother antigens, such as IL-6R molecules from other species. In thecontext of the present invention, multispecific (e.g., bispecific)antibodies that bind to hIL-6R as well as one or more additionalantigens are deemed to “specifically bind” hIL-6R. Moreover, an isolatedantibody may be substantially free of other cellular material and/orchemicals.

Exemplary anti-hIL-6R antibodies that may be included in thepharmaceutical formulations of the present invention are set forth inU.S. Pat. No. 7,582,298, the disclosure of which is incorporated byreference in its entirety.

According to certain embodiments of the present invention, theanti-hIL-6R antibody, or antigen-binding fragment thereof, comprises aheavy chain complementary determining region (HCDR) 1 having an aminoacid sequence selected from the group consisting of SEQ ID NO: 4, 20, 36and 52; a HCDR2 having an amino acid sequence selected from the groupconsisting of SEQ ID NO:6, 22, 38 and 54; and a HCDR3 having an aminoacid sequence selected from the group consisting of SEQ ID NO:2, 18, 34and 50. In certain embodiments, the anti-hIL-6R antibody, orantigen-binding fragment thereof, comprises HCDR1-HCDR2-HCDR3 domains,respectively, selected from the group consisting of: (i) SEQ IDNOs:4-6-8; (ii) SEQ ID NOs:20-22-24; (iii) SEQ ID NOs:36-38-40; and (iv)SEQ ID NOs:52-54-56.

According to certain embodiments of the present invention, theanti-hIL-6R antibody, or antigen-binding fragment thereof, comprises alight chain complementary determining region (LCDR) 1 having an aminoacid sequence selected from the group consisting of SEQ ID NO: 12, 28,44 and 60; a LCDR2 having an amino acid sequence selected from the groupconsisting of SEQ ID NO:14, 30, 46 and 62; and a LCDR3 having an aminoacid sequence selected from the group consisting of SEQ ID NO:16, 32, 48and 64. In certain embodiments, the anti-hIL-6R antibody, orantigen-binding fragment thereof, comprises LCDR1-LCDR2-LCDR3 domains,respectively, selected from the group consisting of: (i) SEQ IDNOs:12-14-16; (ii) SEQ ID NOs:28-30-32; (iii) SEQ ID NOs:44-46-48; and(iv) SEQ ID NOs:60-62-64.

In certain embodiments, the anti-hIL-6R antibody, or antigen-bindingfragment thereof, comprises HCDR1-HCDR2-HCDR3/LCDR1-LCDR2-LCDR3 domains,respectively, selected from the group consisting of: (i) SEQ IDNOs:4-6-8/SEQ ID NOs:12-14-16; (ii) SEQ ID NOs:20-22-24/SEQ IDNOs:28-30-32; (iii) SEQ ID NOs:36-38-40/SEQ ID NOs:44-46-48; and (iv)SEQ ID NOs:52-54-56/SEQ ID NOs:60-62-64.

In certain embodiments, the anti-hIL-6R antibody, or antigen-bindingfragment thereof, comprises a heavy chain variable region (HCVR) havingan amino acid sequence selected from the group consisting of SEQ IDNO:2, 18, 34 and 50. In certain embodiments, the anti-hIL-6R antibody,or antigen-binding fragment thereof, comprises a light chain variableregion (LCVR) having an amino acid sequence selected from the groupconsisting of SEQ ID NO:10, 26, 42 and 58. In certain embodiments, theanti-hIL-6R antibody, or antigen-binding fragment thereof, comprises aHCVR/LCVR amino acid sequence pair selected from the group consisting ofSEQ ID NO:2/10; 18/26; 34/42 and 50/58.

The non-limiting, exemplary antibody used in the Examples herein isreferred to as “mAb1.” This antibody is also referred to in U.S. Pat.No. 7,582,298 as VQ8F11-21. mAb1 (VQ8F11-21) comprises an HCVR/LCVRamino acid sequence pair having SEQ ID NOs:18/26, andHCDR1-HCDR2-HCDR3/LCDR1-LCDR2-LCDR3 domains represented by SEQ IDNOs:20-22-24/SEQ ID NOs:28-30-32.

The amount of antibody, or antigen-binding fragment thereof, containedwithin the pharmaceutical formulations of the present invention may varydepending on the specific properties desired of the formulations, aswell as the particular circumstances and purposes for which theformulations are intended to be used. In certain embodiments, thepharmaceutical formulations may contain about 1 mg/mL to about 500 mg/mLof antibody; about 5 mg/mL to about 400 mg/mL of antibody; about 5 mg/mLto about 200 mg/mL of antibody; about 25 mg/mL to about 180 mg/mL ofantibody; about 25 mg/mL to about 150 mg/mL of antibody; or about 50mg/mL to about 180 mg/mL of antibody. For example, the formulations ofthe present invention may comprise about 1 mg/mL; about 2 mg/mL; about 5mg/mL; about 10 mg/mL; about 15 mg/mL; about 20 mg/mL; about 25 mg/mL;about 30 mg/mL; about 35 mg/mL; about 40 mg/mL; about 45 mg/mL; about 50mg/mL; about 55 mg/mL; about 60 mg/mL; about 65 mg/mL; about 70 mg/mL;about 75 mg/mL; about 80 mg/mL; about 85 mg/mL; about 86 mg/mL; about 87mg/mL; about 88 mg/mL; about 89 mg/mL; about 90 mg/mL; about 95 mg/mL;about 100 mg/mL; about 105 mg/mL; about 110 mg/mL; about 115 mg/mL;about 120 mg/mL; about 125 mg/mL; about 130 mg/mL; about 131 mg/mL;about 132 mg/mL; about 133 mg/mL; about 134 mg/mL; about 135 mg/mL;about 140 mg/mL; about 145 mg/mL; about 150 mg/mL; about 155 mg/mL;about 160 mg/mL; about 165 mg/mL; about 170 mg/mL; about 175 mg/mL;about 180 mg/mL; about 185 mg/mL; about 190 mg/mL; about 195 mg/mL; orabout 200 mg/mL of an antibody or an antigen-binding fragment thereof,that binds specifically to hIL-6R.

EXCIPIENTS and pH

The pharmaceutical formulations of the present invention comprise one ormore excipients. The term “excipient,” as used herein, means anynon-therapeutic agent added to the formulation to provide a desiredconsistency, viscosity or stabilizing effect.

In certain embodiments, the pharmaceutical formulation of the inventioncomprises at least one amino acid. Exemplary amino acids suitable foruse in the formulations of the present invention include, inter alia,arginine and/or histidine.

The amount of amino acid contained within the pharmaceuticalformulations of the present invention may vary depending on the specificproperties desired of the formulations, as well as the particularcircumstances and purposes for which the formulations are intended to beused. In certain embodiments, the formulations may contain about 1 mM toabout 200 mM of an amino acid; about 2 mM to about 100 mM of an aminoacid; about 5 mM to about 50 mM of an amino acid; or about 10 mM toabout 25 mM of an amino acid. For example, the pharmaceuticalformulations of the present invention may comprise about 1 mM; about 1.5mM; about 2 mM; about 2.5 mM; about 3 mM; about 3.5 mM; about 4 mM;about 4.5 mM; about 5 mM; about 5.5 mM; about 6 mM; about 6.5 mM; about7 mM; about 7.5 mM; about 8 mM; about 8.5 mM; about 9 mM; about 9.5 mM;about 10 mM; about 10.5 mM; about 11 mM; about 11.5 mM; about 12 mM;about 12.5 mM; about 13 mM; about 13.5 mM; about 14 mM; about 14.5 mM;about 15 mM; about 15.5 mM; 16 mM; about 16.5 mM; about 17 mM; about17.5 mM; about 18 mM; about 18.5 mM; about 19 mM; about 19.5 mM; about20 mM; about 20.5 mM; about 21 mM; about 21.5 mM; about 22 mM; about22.5 mM; about 23 mM; about 23.5 mM; about 24 mM; about 24.5 mM; about25 mM; about 25.5 mM; about 26 mM; about 26.5 mM; about 27 mM; about27.5 mM; about 28 mM; about 28.5 mM; about 29 mM; about 29.5 mM; about30 mM; about 35 mM; about 40 mM; about 45 mM; or about 50 mM of an aminoacid (e.g., histidine and/or arginine).

The pharmaceutical formulations of the present invention may alsocomprise one or more carbohydrate, e.g., one or more sugar. The sugarcan be a reducing sugar or a non-reducing sugar. “Reducing sugars”include, e.g., sugars with a ketone or aldehyde group and contain areactive hemiacetal group, which allows the sugar to act as a reducingagent. Specific examples of reducing sugars include fructose, glucose,glyceraldehyde, lactose, arabinose, mannose, xylose, ribose, rhamnose,galactose and maltose. Non-reducing sugars can comprise an anomericcarbon that is an acetal and is not substantially reactive with aminoacids or polypeptides to initiate a Maillard reaction. Specific examplesof non-reducing sugars include sucrose, trehalose, sorbose, sucralose,melezitose and raffinose. Sugar acids include, for example, saccharicacids, gluconate and other polyhydroxy sugars and salts thereof.

The amount of sugar contained within the pharmaceutical formulations ofthe present invention will vary depending on the specific circumstancesand intended purposes for which the formulations are used. In certainembodiments, the formulations may contain about 0.1% to about 20% sugar;about 0.5% to about 20% sugar; about 1% to about 20% sugar; about 2% toabout 15% sugar; about 3% to about 10% sugar; about 4% to about 10%sugar; or about 5% to about 10% sugar. For example, the pharmaceuticalformulations of the present invention may comprise about 0.5%; about1.0%; about 1.5%; about 2.0%; about 2.5%; about 3.0%; about 3.5%; about4.0%; about 4.5%; about 5.0%; about 5.5%; about 6.0%; 6.5%; about 7.0%;about 7.5%; about 8.0%; about 8.5%; about 9.0%; about 9.5%; about 10.0%;about 10.5%; about 11.0%; about 11.5%; about 12.0%; about 12.5%; about13.0%; about 13.5%; about 14.0%; about 14.5%; about 15.0%; about 15.5%;about 16.0%; 16.5%; about 17.0%; about 17.5%; about 18.0%; about 18.5%;about 19.0%; about 19.5%; or about 20.0% sugar (e.g., sucrose).

The pharmaceutical formulations of the present invention may alsocomprise one or more surfactant. As used herein, the term “surfactant”means a substance which reduces the surface tension of a fluid in whichit is dissolved and/or reduces the interfacial tension between oil andwater. Surfactants can be ionic or non-ionic. Exemplary non-ionicsurfactants that can be included in the formulations of the presentinvention include, e.g., alkyl poly(ethylene oxide), alkylpolyglucosides (e.g., octyl glucoside and decyl maltoside), fattyalcohols such as cetyl alcohol and oleyl alcohol, cocamide MEA, cocamideDEA, and cocamide TEA. Specific non-ionic surfactants that can beincluded in the formulations of the present invention include, e.g.,polysorbates such as polysorbate 20, polysorbate 28, polysorbate 40,polysorbate 60, polysorbate 65, polysorbate 80, polysorbate 81, andpolysorbate 85; poloxamers such as poloxamer 188, poloxamer 407;polyethylene-polypropylene glycol; or polyethylene glycol (PEG).Polysorbate 20 is also known as TWEEN 20, sorbitan monolaurate andpolyoxyethylenesorbitan monolaurate.

The amount of surfactant contained within the pharmaceuticalformulations of the present invention may vary depending on the specificproperties desired of the formulations, as well as the particularcircumstances and purposes for which the formulations are intended to beused. In certain embodiments, the formulations may contain about 0.05%to about 5% surfactant; or about 0.1% to about 0.2% surfactant. Forexample, the formulations of the present invention may comprise about0.05%; about 0.06%; about 0.07%; about 0.08%; about 0.09%; about 0.10%;about 0.11%; about 0.12%; about 0.13%; about 0.14%; about 0.15%; about0.16%; about 0.17%; about 0.18%; about 0.19%; about 0.20%; about 0.21%;about 0.22%; about 0.23%; about 0.24%; about 0.25%; about 0.26%; about0.27%; about 0.28%; about 0.29%; or about 0.30% surfactant (e.g.,polysorbate 20).

The pharmaceutical formulations of the present invention may have a pHof from about 5.0 to about 8.0. For example, the formulations of thepresent invention may have a pH of about 5.0; about 5.2; about 5.4;about 5.6; about 5.8; about 6.0; about 6.2; about 6.4; about 6.6; about6.8; about 7.0; about 7.2; about 7.4; about 7.6; about 7.8; or about8.0.

EXEMPLARY FORMULATIONS

According to one aspect of the present invention, the pharmaceuticalformulation comprises: (i) a human antibody that specifically binds tohIL-6R (e.g., mAb1); (ii) an amino acid (e.g., histidine); and (iii) asugar (e.g., sucrose). Specific, non-limiting exemplary embodimentsencompassed by this aspect of the invention are set forth in Table 1.

TABLE 1 Exemplary Pharmaceutical Formulations Comprising mAb1, Histidineand Sucrose mAb1 (mg/ml) 25 50 100 150 25 50 100 150 25 50 100 150 25 50100 150 histidine (mM) 10 10 10 10 10 10 10 10 10 10 10 10 10 10 10 10sucrose (%) 1 1 1 1 2 2 2 2 4 4 4 4 6 6 6 6

According to another aspect of the present invention, the pharmaceuticalformulation comprises: (i) a human antibody that specifically binds tohIL-6R (e.g., mAb1); (ii) an amino acid (e.g., histidine); (iii) a sugar(e.g., sucrose); and (iv) a surfactant (e.g., polysorbate 20). Specific,non-limiting exemplary embodiments encompassed by this aspect of theinvention are set forth in Tables 2A and 2B.

TABLE 2A Exemplary Pharmaceutical Formulations Comprising mAb1,Histidine, Sucrose and Polysorbate 20 mAb1 (mg/ml) 25 50 100 150 25 50100 150 25 50 100 150 histidine (mM) 10 10 10 10 10 10 10 10 10 10 10 10sucrose (%) 2 2 2 2 5 5 5 5 10 10 10 10 polysorbate 20 (%) 0.1 0.1 0.10.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1

TABLE 2B Exemplary Pharmaceutical Formulations Comprising mAb1,Histidine, Sucrose and Polysorbate 20 mAb1 (mg/ml) 25 50 100 150 25 50100 150 25 50 100 150 histidine (mM) 10 10 10 10 10 10 10 10 10 10 10 10sucrose (%) 2 2 2 2 5 5 5 5 10 10 10 10 polysorbate 20 (%) 0.2 0.2 0.20.2 0.2 0.2 0.2 0.2 0.2 0.2 0.2 0.2

According to another aspect of the present invention, the pharmaceuticalformulation comprises: (i) a human antibody that specifically binds tohIL-6R (e.g., mAb1); (ii) a first amino acid (e.g., histidine); (iii) asugar (e.g., sucrose); (iv) a surfactant (e.g., polysorbate 20); and (v)a second amino acid (e.g., arginine). Specific, non-limiting exemplaryembodiments encompassed by this aspect of the invention are set forth inTables 3A, 3B, 3C, 3D, 3E and 3F.

TABLE 3A Exemplary Pharmaceutical Formulations Comprising mAb1,Histidine, Sucrose, Polysorbate 20 and Arginine mAb1 (mg/ml) 25 50 100150 25 50 100 150 25 50 100 150 histidine (mM) 10 10 10 10 10 10 10 1010 10 10 10 sucrose (%) 2 2 2 2 5 5 5 5 10 10 10 10 polysorbate 20 (%)0.2 0.2 0.2 0.2 0.2 0.2 0.2 0.2 0.2 0.2 0.2 0.2 arginine (mM) 10 10 1010 10 10 10 10 10 10 10 10

TABLE 3B Exemplary Pharmaceutical Formulations Comprising mAb1,Histidine, Sucrose, Polysorbate 20 and Arginine mAb1 (mg/ml) 25 50 100150 25 50 100 150 25 50 100 150 histidine (mM) 10 10 10 10 10 10 10 1010 10 10 10 sucrose (%) 2 2 2 2 5 5 5 5 10 10 10 10 polysorbate 20 (%)0.2 0.2 0.2 0.2 0.2 0.2 0.2 0.2 0.2 0.2 0.2 0.2 arginine (mM) 25 25 2525 25 25 25 25 25 25 25 25

TABLE 3C Exemplary Pharmaceutical Formulations Comprising mAb1,Histidine, Sucrose, Polysorbate 20 and Arginine mAb1 (mg/ml) 25 50 100150 25 50 100 150 25 50 100 150 histidine (mM) 25 25 25 25 25 25 25 2525 25 25 25 sucrose (%) 2 2 2 2 5 5 5 5 10 10 10 10 polysorbate 20 (%)0.2 0.2 0.2 0.2 0.2 0.2 0.2 0.2 0.2 0.2 0.2 0.2 arginine (mM) 10 10 1010 10 10 10 10 10 10 10 10

TABLE 3D Exemplary Pharmaceutical Formulations Comprising mAb1,Histidine, Sucrose, Polysorbate 20 and Arginine mAb1 (mg/ml) 25 50 100150 25 50 100 150 25 50 100 150 histidine (mM) 25 25 25 25 25 25 25 2525 25 25 25 sucrose (%) 2 2 2 2 5 5 5 5 10 10 10 10 polysorbate 20 (%)0.2 0.2 0.2 0.2 0.2 0.2 0.2 0.2 0.2 0.2 0.2 0.2 arginine (mM) 25 25 2525 25 25 25 25 25 25 25 25

TABLE 3E Exemplary Pharmaceutical Formulations Comprising mAb1,Histidine, Sucrose, Polysorbate 20 and Arginine mAb1 (mg/ml) 25 50 100150 25 50 100 150 25 50 100 150 histidine (mM) 25 25 25 25 25 25 25 2525 25 25 25 sucrose (%) 2 2 2 2 5 5 5 5 10 10 10 10 polysorbate 20 (%)0.2 0.2 0.2 0.2 0.2 0.2 0.2 0.2 0.2 0.2 0.2 0.2 arginine (mM) 50 50 5050 50 50 50 50 50 50 50 50

TABLE 3F Exemplary Pharmaceutical Formulations Comprising mAb1,Histidine, Sucrose, Polysorbate 20 and Arginine mAb1 (mg/ml) 160 170 175180 160 170 175 180 160 170 175 180 histidine (mM) 25 25 25 25 25 25 2525 25 25 25 25 sucrose (%) 2 2 2 2 5 5 5 5 10 10 10 10 polysorbate 20(%) 0.2 0.2 0.2 0.2 0.2 0.2 0.2 0.2 0.2 0.2 0.2 0.2 arginine (mM) 50 5050 50 50 50 50 50 50 50 50 50

Additional non-limiting examples of pharmaceutical formulationsencompassed by the present invention are set forth elsewhere herein,including the working Examples presented below.

STABILITY AND VISCOSITY OF THE PHARMACEUTICAL FORMULATIONS

The pharmaceutical formulations of the present invention typicallyexhibit high levels of stability. The term “stable,” as used herein inreference to the pharmaceutical formulations, means that the antibodieswithin the pharmaceutical formulations retain an acceptable degree ofstructure and/or function and/or biological activity after storage for adefined amount of time. A formulation may be stable even though theantibody contained therein does not maintain 100% of its structureand/or function and/or biological activity after storage for a definedamount of time. Under certain circumstances, maintenance of about 80%,about 85%, about 90%, about 95%, about 96%, about 97%, about 98% orabout 99% of an antibody's structure and/or function and/or biologicalactivity after storage for a defined amount of time may be regarded as“stable.”

Stability can be measured, inter alia, by determining the percentage ofnative antibody remaining in the formulation after storage for a definedamount of time at a given temperature. The percentage of native antibodycan be determined by, inter alia, size exclusion chromatography (e.g.,size exclusion high performance liquid chromatography [SE-HPLC]). An“acceptable degree of stability,” as that phrase is used herein, meansthat at least 90% of the native form of the antibody can be detected inthe formulation after storage for a defined amount of time at a giventemperature. In certain embodiments, at least about 90%, 91%, 92%, 93%,94%, 95%, 96%, 97%, 98%, 99% or 100% of the native form of the antibodycan be detected in the formulation after storage for a defined amount oftime at a given temperature. The defined amount of time after whichstability is measured can be at least 1 month, at least 2 months, atleast 3 months, at least 4 months, at least 5 months, at least 6 months,at least 7 months, at least 8 months, at least 9 months, at least 10months, at least 11 months, at least 12 months, at least 18 months, atleast 24 months, or more. The temperature at which the pharmaceuticalformulation may be stored when assessing stability can be anytemperature from about −80° C. to about 45° C., e.g., storage at about−30° C., about −20° C., about 0° C., about 5° C., about 25° C., or about45° C. For example, a pharmaceutical formulation may be deemed stable ifafter 3 months of storage at 5° C., greater than about 90%, 95%, 96% or97% of native antibody is detected by SE-HPLC. A pharmaceuticalformulation may also be deemed stable if after 6 months of storage at 5°C., greater than about 90%, 95%, 96% or 97% of native antibody isdetected by SE-HPLC. A pharmaceutical formulation may also be deemedstable if after 9 months of storage at 5° C., greater than about 90%,95%, 96% or 97% of native antibody is detected by SE-HPLC. Apharmaceutical formulation may also be deemed stable if after 3 monthsof storage at 25° C., greater than about 90%, 95%, 96% or 97% of nativeantibody is detected by SE-HPLC. A pharmaceutical formulation may alsobe deemed stable if after 6 months of storage at 25° C., greater thanabout 90%, 95%, 96% or 97% of native antibody is detected by SE-HPLC. Apharmaceutical formulation may also be deemed stable if after 9 monthsof storage at 25° C., greater than about 90%, 95%, 96% or 97% of nativeantibody is detected by SE-HPLC.

Other methods may be used to assess the stability of the formulations ofthe present invention such as, e.g., differential scanning calorimetry(DSC) to determine thermal stability, controlled agitation to determinemechanical stability, and absorbance at about 350 nm or about 405 nm todetermine solution turbidities. For example, a formulation of thepresent invention may be considered stable if, after 6 or more months ofstorage at about 5° C. to about 25° C., the change in OD405 of theformulation is less than about 0.05 (e.g., 0.04, 0.03, 0.02, 0.01, orless) from the OD405 of the formulation at t=0.

Stability may also be assessed by measuring the biological activityand/or binding affinity of the antibody to its target. For example, aformulation of the present invention may be regarded as stable if, afterstorage at e.g., 5° C., 25° C., 45° C., etc. for a defined amount oftime (e.g., 1 to 12 months), the anti-IL-6R antibody contained withinthe formulation binds to IL-6R with an affinity that is at least 50%,60%, 70%, 80%, 90%, 95%, or more of the binding affinity of the antibodyprior to said storage. Additional methods for assessing the stability ofan antibody in formulation are demonstrated in the Examples presentedbelow.

In the fluid form, the pharmaceutical formulations of the presentinvention may, in certain embodiments, exhibit low to moderate levels ofviscosity. “Viscosity” as used herein may be “kinematic viscosity” or“absolute viscosity.” “Kinematic viscosity” is a measure of theresistive flow of a fluid under the influence of gravity. When twofluids of equal volume are placed in identical capillary viscometers andallowed to flow by gravity, a viscous fluid takes longer than a lessviscous fluid to flow through the capillary. For example, if one fluidtakes 200 seconds to complete its flow and another fluid takes 400seconds, the second fluid is twice as viscous as the first on akinematic viscosity scale. “Absolute viscosity”, sometimes calleddynamic or simple viscosity, is the product of kinematic viscosity andfluid density (Absolute Viscosity=Kinematic Viscosity x Density). Thedimension of kinematic viscosity is L²/T where L is a length and T is atime. Commonly, kinematic viscosity is expressed in centistokes (cSt).The SI unit of kinematic viscosity is mm²/s, which is 1 cSt. Absoluteviscosity is expressed in units of centipoise (cP). The SI unit ofabsolute viscosity is the milliPascal-second (mPa·s), where 1 cP=1mPa·s.

As used herein, a low level of viscosity, in reference to a fluidformulation of the present invention, will exhibit an absolute viscosityof less than about 20 cPoise (cP). For example, a fluid formulation ofthe invention will be deemed to have “low viscosity,” if, when measuredusing standard viscosity measurement techniques, the formulationexhibits an absolute viscosity of about 19 cP, about 18 cP, about 17 cP,about 16 cP, about 15 cP, about 14 cP, about 13 cP, about 12 cP, about11 cP, about 10 cP, about 9 cP, about 8 cP, about 7 cP, about 6 cP,about 5 cP, about 4 cP, or less. As used herein, a moderate level ofviscosity, in reference to a fluid formulation of the present invention,will exhibit an absolute viscosity of between about 30 cP and about 20cP. For example, a fluid formulation of the invention will be deemed tohave “moderate viscosity,” if when measured using standard viscositymeasurement techniques, the formulation exhibits an absolute viscosityof about 30 cP, about 29 cP, about 28 cP, about 27 cP, about 26 cP,about 25 cP, about 24 cP, about 23 cP, about 22 cP, about 21 cP or about20 cP.

As illustrated in Example 6 below, the present inventors have made thesurprising discovery that low to moderate viscosity fluid formulationscomprising high concentrations of an anti-hIL-6R antibody (e.g., up toat least 175 mg/mL) can be obtained by formulating the antibody with 25mM to 100 mM histidine and 25 mM to 50 mM arginine. In addition, it wasfurther discovered that the viscosity of the formulation could bedecreased to an even greater extent by adjusting the sucrose content toless than 10%.

CONTAINERS FOR THE PHARMACEUTICAL FORMULATIONS AND METHODS OFADMINISTRATION

The pharmaceutical formulations of the present invention may becontained within any container suitable for storage of medicines andother therapeutic compositions. For example, the pharmaceuticalformulations may be contained within a sealed and sterilized plastic orglass container having a defined volume such as a vial, ampule, syringe,cartridge, or bottle. Different types of vials can be used to containthe formulations of the present invention including, e.g., clear andopaque (e.g., amber) glass or plastic vials. Likewise, any type ofsyringe can be used to contain and/or administer the pharmaceuticalformulations of the present invention.

The pharmaceutical formulations of the present invention may becontained within “normal tungsten” syringes or “low tungsten” syringes.As will be appreciated by persons of ordinary skill in the art, theprocess of making glass syringes generally involves the use of a hottungsten rod which functions to pierce the glass thereby creating a holefrom which liquids can be drawn and expelled from the syringe. Thisprocess results in the deposition of trace amounts of tungsten on theinterior surface of the syringe. Subsequent washing and other processingsteps can be used to reduce the amount of tungsten in the syringe. Asused herein, the term “normal tungsten” means that the syringe containsgreater than 500 parts per billion (ppb) of tungsten. The term “lowtungsten” means that the syringe contains less than 500 ppb of tungsten.For example, a low tungsten syringe, according to the present invention,can contain less than about 490, 480, 470, 460, 450, 440, 430, 420, 410,390, 350, 300, 250, 200, 150, 100, 90, 80, 70, 60, 50, 40, 30, 20, 10 orfewer ppb of tungsten.

The rubber plungers used in syringes, and the rubber stoppers used toclose the openings of vials, may be coated to prevent contamination ofthe medicinal contents of the syringe or vial and/or to preserve theirstability. Thus, pharmaceutical formulations of the present invention,according to certain embodiments, may be contained within a syringe thatcomprises a coated plunger, or within a vial that is sealed with acoated rubber stopper. For example, the plunger or stopper may be coatedwith a fluorocarbon film. Examples of coated stoppers and/or plungerssuitable for use with vials and syringes containing the pharmaceuticalformulations of the present invention are mentioned in, e.g., U.S. Pat.Nos. 4,997,423; 5,908,686; 6,286,699; 6,645,635; and 7,226,554, thecontents of which are incorporated by reference herein in theirentireties. Particular exemplary coated rubber stoppers and plungersthat can be used in the context of the present invention arecommercially available under the tradename “FluroTec®,” available fromWest Pharmaceutical Services, Inc. (Lionville, Pa.).

According to certain embodiments of the present invention, thepharmaceutical formulations may be contained within a low tungstensyringe that comprises a fluorocarbon- coated plunger. As discussed inthe Examples section below, the combination of a low tungsten syringeand a fluorocarbon-coated plunger was observed to yield surprisingstability characteristics with regard to the pharmaceutical formulationsof the present invention.

The pharmaceutical formulations can be administered to a patient byparenteral routes such as injection (e.g., subcutaneous, intravenous,intramuscular, intraperitoneal, etc.) or percutaneous, mucosal, nasal,pulmonary and/or oral administration. Numerous reusable pen and/orautoinjector delivery devices can be used to subcutaneously deliver thepharmaceutical formulations of the present invention. Examples include,but are not limited to AUTOPEN™ (Owen Mumford, Inc., Woodstock, UK),DISETRONIC™ pen (Disetronic Medical Systems, Bergdorf, Switzerland),HUMALOG MIX 75/25™ pen, HUMALOG™pen, HUMALIN 70/30™ pen (Eli Lilly andCo., Indianapolis, Ind.), NOVOPEN™ I, II and III (Novo Nordisk,Copenhagen, Denmark), NOVOPEN JUNIORT™ Novo Nordisk, Copenhagen,Denmark), BD™ pen (Becton Dickinson, Franklin Lakes, N.J.), OPTIPEN™,OPTIPEN PRO™, OPTIPEN STARLET™, and OPTICLIK™ (sanofi-aventis,Frankfurt, Germany), to name only a few. Examples of disposable penand/or autoinjector delivery devices having applications in subcutaneousdelivery of a pharmaceutical composition of the present inventioninclude, but are not limited to the SOLOSTAR™ pen (sanofi-aventis), theFLEXPEN™ (Novo Nordisk), and the KWIKPEN™ (Eli Lilly), the SURECLICK™Autoinjector (Amgen, Thousand Oaks, Calif.), the PENLET™ (Haselmeier,Stuttgart, Germany), the EPIPEN (Dey, L.P.), and the HUMIRA™ Pen (AbbottLabs, Abbott Park, Ill.), to name only a few.

The use of a microinfusor to deliver the pharmaceutical formulations ofthe present invention is also contemplated herein. As used herein, theterm “microinfusor” means a subcutaneous delivery device designed toslowly administer large volumes (e.g., up to about 2.5 mL or more) of atherapeutic formulation over a prolonged period of time (e.g., about 10,15, 20, 25, 30 or more minutes). See, e.g., U.S. Pat. No. 6,629,949;U.S. Pat. No. 6,659,982; and Meehan et al., J. Controlled Release46:107-116 (1996). Microinfusors are particularly useful for thedelivery of large doses of therapeutic proteins contained within highconcentration (e.g., about 100, 125, 150, 175, 200 or more mg/mL) and/orviscous solutions.

THERAPEUTIC USES OF THE PHARMACEUTICAL FORMULATIONS

The pharmaceutical formulations of the present invention are useful,inter alia, for the treatment, prevention and/or amelioration of anydisease or disorder associated withIL-6 activity, including diseases ordisorders mediated by activation of the IL-6 receptor. Exemplary,non-limiting diseases and disorders that can be treated and/or preventedby the administration of the pharmaceutical formulations of the presentinvention include, e.g., rheumatoid arthritis, ankylosing spondylitis,Crohn's disease, ulcerative colitis, pancreatitis, juvenile idiopathicarthritis, vasculitis, Kawasaki disease, systemic lupus erythematosis,psoriasis, psoriatic arthritis, Sjogren syndrome, Still's disease,Castleman's disease, multiple sclerosis, diseases associated withabnormal blood coagulation or fibrinolysis (e.g., thrombosis), cancer(e.g., breast cancer, leukemia, ovarian cancer, melanoma, prostatecancer, pancreatic cancer, lymphoma, lung cancer, renal cell carcinoma,colorectal cancer, multiple myeloma, etc.), cachexia, chronic rejectionof transplanted organs and cells, cardiopathy, viral infection (e.g.,HIV infection, EBV infection, etc.), plasmacytosis,hyperimmunoglobulinemia, anemia, nephritis, mesothelioma, and hearingloss and other inner ear disorders.

Thus, the present invention includes methods of treating, preventing,and/or ameliorating any disease or disorder associated withIL-6 activityor IL-6R activation (including any of the above mentioned exemplarydiseases, disorders and conditions). The therapeutic methods of thepresent invention comprise administering to a subject any formulationcomprising an anti-hIL-6R antibody as disclosed herein. The subject towhich the pharmaceutical formulation is administered can be, e.g., anyhuman or non-human animal that is in need of such treatment, preventionand/or amelioration, or who would otherwise benefit from the inhibitionor attenuation of IL-6 and/or IL-6R-mediated activity. For example, thesubject can be an individual that is diagnosed with, or who is deemed tobe at risk of being afflicted by any of the aforementioned diseases ordisorders. The present invention further includes the use of any of thepharmaceutical formulations disclosed herein in the manufacture of amedicament for the treatment, prevention and/or amelioration of anydisease or disorder associated withIL-6 activity or IL-6R activation(including any of the above mentioned exemplary diseases, disorders andconditions).

EXAMPLES

The following examples are put forth so as to provide those of ordinaryskill in the art with a complete disclosure and description of how tomake and use the methods and compositions of the invention, and are notintended to limit the scope of what the inventors regard as theirinvention. Efforts have been made to ensure accuracy with respect tonumbers used (e.g., amounts, temperature, etc.) but some experimentalerrors and deviations should be accounted for. Unless indicatedotherwise, parts are parts by weight, molecular weight is averagemolecular weight, temperature is in degrees Centigrade, and pressure isat or near atmospheric.

Example 1 Stability of a Fully Human Anti-Human Interleukin-6 Receptor(IL-6R) Antibody (“mAb1”) After Storage at Low Temperatures

In this Example, various formulations were created containing ananti-human IL-6R antibody without excipients. The exemplary antibodyused in this and all subsequent Examples set forth below is an antibodycomprising a heavy chain variable region (HCVR) with the amino acidsequence of SEQ ID NO:18, and a light chain variable region (LCVR) withthe amino acid sequence of SEQ ID NO:26. This antibody is referred toherein as “mAb1”.

As a preliminary experiment, the stability of mAb1 in liquid solutionwas determined following various amounts of time in frozen storage at−20° C., −30° C. and −80° C. The concentration of mAb1 used in thisExample was 128 mg/mL. At various time points, the stability of mAb1 wasdetermined by size exclusion high performance liquid chromatography(SE-HPLC) and by cation exchange high performance liquid chromatography(CEX-HPLC). Stability was assessed based on the percentage of nativemAb1 remaining in the sample (by SE-HPLC; Table 4) and by the percentageof acidic species observed in the sample (by CEX-HPLC; Table 5) (Anincrease in percent acidic species is consistent with deamidation of theantibody and is thus considered an undesired phenomenon with respect tothe pharmaceutical formulations of the present invention).

The results of Tables 3 and 4 are depicted in FIGS. 1 and 2,respectively. These results show that mAb1 can remain stable at aconcentration of 128 mg/mL for at least 9 months when stored at −80° C.

TABLE 4 % Native mAb1 Remaining (SE-HPLC) Time Storage Temperature(months) −80° C. −30° C. −20° C. 0 95.9 95.9 95.9 1 95.7 94.3 93.2 395.6 93.6 89.3 6 96.1 96.0 88.9 9 95.6 91.6 87.9

TABLE 5 % Acidic Species (CEX-HPLC) Time Storage Temperature (months)−80° C. −30° C. −20° C. 0 28.6 28.6 28.6 1 27.3 28.0 28.1 3 27.3 27.728.3 6 28.6 29.6 29.3 9 29.0 28.8 29.0

Example 2 Stability of mAb1 Formulations Containing Minimal Excipients

Eight different formulations containing mAb1 and minimal excipients asshown in Table 6 were prepared.

TABLE 6 mAb1 Minimal Excipient Formulations Formulation Excipient mAb1(mg/mL) 1 0.13% polysorbate 20 80 6% sucrose 2 0.13% polysorbate 20 80 31% sucrose 80 4 2% sucrose 80 5 4% sucrose 80 6 6% sucrose 80 7 none 808 none 65 All formulations contain 10 mM histidine, pH 6.0

The formulations were tested for stability by SE-HPLC after variousamounts of time at −30° C. and −20° C. The results, expressed in percentof native mAb1 remaining, are shown in Tables 7 (−30° C. storage) and 8(−20° C.).

TABLE 7 % Native mAb1 Remaining (SE-HPLC) After Storage at −30° C. TimeFormulation # (see Table 6) (months) 1 2 3 4 5 6 7 8 0 96.4 96.2 96.496.5 96.5 96.6 96.2 96.5 1 96.4 95.1 96.3 96.3 96.4 96.6 95.1 95.3 296.4 94.7 96.0 96.4 96.5 96.0 95.0 95.4 3 96.5 94.7 96.3 96.7 96.7 96.794.4 94.9 4 97.2 95.2 96.7 97.4 97.3 97.3 95.1 95.7 6 97.0 94.2 96.296.8 97.1 96.9 94.0 94.5 9 96.7 93.6 96.0 96.6 96.5 96.9 93.4 93.8

TABLE 8 % Native mAb1 Remaining (SE-HPLC) After Storage at −20° C. TimeFormulation # (see Table 6) (months) 1 2 3 4 5 6 7 8 0 96.4 96.2 96.496.5 96.5 96.6 96.2 96.5 1 96.7 94.5 96.0 96.8 96.5 96.3 94.1 94.5 296.4 90.9 95.3 96.4 96.4 96.4 90.9 91.8 3 96.9 90.1 95.1 96.6 96.6 96.790.5 90.9 4 97.2 90.7 95.8 97.4 97.1 97.1 91.4 91.8 6 96.9 86.9 94.196.9 96.9 97.1 87.5 88.5 9 96.5 86.0 93.3 96.6 96.6 96.7 86.7 87.5

The results of Tables 7 and 8 are depicted in FIGS. 3 and 4,respectively. As shown in this Example, the stability of mAb1 wasmaintained to a significant extent in formulations 1, 4, 5 and 6 afterseveral months of storage at −20° C. and −30° C. These results indicatethat the stability of mAb1 at −20° C. and −30° C. can be enhanced by theaddition of at least 2% sucrose.

Example 3 Stabilized Formulation of mAbi

A stabilized formulation containing various concentrations of mAb1 wasprepared for use in Examples 4 and 5 below. This formulation, designated“Formulation A”, is shown in Table 9.

TABLE 9 Stabilized mAb1 Formulation “A” Component Formulation A mAb125-100 mg/mL Histidine 10 mM Polysorbate 20 0.2% Sucrose  10% pHadjusted to 6.0

Example 4 Stability of Formulation A After Storage at 5° C.

Formulation A (see Example 3) containing 25, 50 or 100 mg/mL mAb1 wastested for stability after several months of storage at 5° C. in clearvials. Stability was assessed by the following parameters: (a) visualappearance; (b) turbidity (OD 405 nm); (c) pH; (d) percent total mAb1recovered (as measured by RP-HPLC); (d) percent native mAb1 recovered(as measured by SE-HPLC); (e) percent main peak mAb1 recovered (asmeasured by CEX-HPLC); and (f) percent acidic species mAb1 recovered (asmeasured by CEX-HPLC). The stability results for Formulation Acontaining 25, 50 and 100 mg/mL of mAb1 are summarized in Tables 10, 11and 12, respectively.

TABLE 10 Stability of Formulation A Containing 25 mg/mL mAb1 AfterStorage at 5° C. in Clear Vials Length of 5° C. Storage (months)Parameter 0 1 2 3 6 9 12 Visual Pass Pass Pass Pass Pass Pass PassAppearance Turbidity 0.00 0.00 0.00 0.00 0.00 0.00 0.00 (OD 405 nm) pH6.0 6.1 6.1 6.1 6.1 6.1 6.1 % Total mAb1 100 99 101 112 103 94 101Recovered % Native mAb1 97.5 98.0 97.5 97.6 97.5 97.5 97.8 Recovered %Main Peak 58.4 57.9 58.7 58.1 57.9 57.9 58.4 mAb1 Recovered % AcidicSpecies 26.5 28.0 26.5 28.0 27.3 28.0 27.9 mAb1 Recovered

TABLE 11A Stability of Formulation A Containing 50 mg/mL mAb1 AfterStorage at 5° C. in Clear Vials (0-9 months Length of 5° C. Storage(months) Parameter 0 1 2 3 6 9 Visual Pass Pass Pass Pass Pass PassAppearance Turbidity 0.00 0.00 0.00 0.00 0.00 0.00 (OD 405 nm) pH 5.85.9 5.8 5.9 5.9 6.0 % Total mAb1 100 99 104 106 100 109 Recovered %Native mAb1 97.4 97.5 97.3 97.2 97.3 97.2 Recovered % Main Peak 57.156.7 58.0 54.2 53.3 57.9 mAb1 Recovered % Acidic Species 27.6 26.7 27.628.5 26.8 26.9 mAb1 Recovered

TABLE 11B Stability of Formulation A Containing 50 mg/mL mAb1 AfterStorage at 5° C. in Clear Vials (12-24 months) Length of 5° C. Storage(months) Parameter 12 18 24 Visual Appearance Pass Pass Pass Turbidity(OD 405 nm) 0.00 0.00 0.00 pH 5.9 6.0 5.9 % Total mAb1 103 107 105Recovered % Native mAb1 97.1 97.1 96.9 Recovered % Main Peak mAb1 56.457.1 56.4 Recovered % Acidic Species 28.1 28.3 29.0 mAb1 Recovered

TABLE 12A Stability of Formulation A Containing 100 mg/mL mAb1 AfterStorage at 5° C. in Clear Vials (0-9 months) Length of 5° C. Storage(months) Parameter 0 1 2 3 6 9 Visual Pass Pass Pass Pass Pass PassAppearance Turbidity 0.00 0.00 0.00 0.00 0.00 0.00 (OD 405 nm) pH 5.96.0 5.9 5.9 5.9 5.9 % Total mAb1 100 99 100 107 101 106 Recovered %Native mAb1 97.3 97.0 97.0 97.1 96.9 96.9 Recovered % Main Peak 55.155.5 57.9 55.9 55.4 56.8 mAb1 Recovered % Acidic Species 27.6 26.9 27.429.6 27.4 27.4 mAb1 Recovered

TABLE 12B Stability of Formulation A Containing 100 mg/mL mAb1 AfterStorage at 5° C. in Clear Vials (12-24 months) Length of 5° C. Storage(months) Parameter 12 18 24 Visual Appearance Pass Pass Pass Turbidity(OD 405 nm) 0.00 0.00 0.00 pH 6.0 6.0 6.0 % Total mAb1 101 102 103Recovered % Native mAb1 96.8 96.7 96.5 Recovered % Main Peak mAb1 57.357.5 56.7 Recovered % Acidic Species 27.3 28.1 28.7 mAb1 Recovered

The results of this Example demonstrate that Formulation A containing25, 50 or 100 mg/mL mAb1 remained stable after at least 9 months ofstorage, at 5° C. in clear vials, with about 97% or more of native mAb1remaining in all samples after 9 months of storage under suchconditions. For the 50 and 100 mg/mL formulations, 96.9% and 96.5% ofnative mAb1, respectively, was detected after up to 24 months of storageat 5° C. In addition, the percent acidic species remained at 29% orlower for all time points analyzed, thus confirming the stability of theformulations.

Similar stability studies were also carried out using Formulation Acontaining 75 mg/mL mAb1 following storage at 2-8° C. No significantdegradation was observed for any of the concentrations tested after 24months of 2-8° C. storage as determined by SE-HPLC and CEX-HPLC (datanot shown).

Example 5 Stability of Formulation A Manufactured in Clear and AmberGlass Vials

Additional experiments were conducted to compare the stability ofFormulation A (see Example 3) containing 25 and 100 mg/mL mAb1manufactured in amber glass vials to the same formulation manufacturedin clear vials. Two types of amber vials were used in this Example: 5 mLand 20 mL amber vials. Stability was assessed following storage at 5°C., 25° C. or 45° C. based on the same parameters as used in Example 4.The results for the 25 mg/mL and 100 mg/mL formulations are summarizedin Tables 13 through 21.

TABLE 13 Stability of Formulation A Containing 25 mg/mL mAb1 AfterStorage at 5° C. in 5 mL clear Vials t = 0 Storage at 5° C. (months)Parameter 0 1 2 3 6 9 12 Visual Appearance Pass Pass Pass Pass Pass PassPass Turbidity 0.0 0.0 0.0 0.0 0.0 0.0 0.0 (OD 405 nm) pH 6.0 6.0 6.06.0 6.0 6.0 6.1 % Total mAb1 100 106 103 98 100 100 101 Recovered %Native mAb1 97.1 97.0 96.9 96.9 97.0 96.4 96.6 Recovered % Main Peak57.8 56.8 56.2 54.2 56.4 56.4 56.8 mAb1 Recovered % Acidic Species 27.930.7 30.8 33.0 30.6 29.8 30.1 mAb1 Recovered

TABLE 14 Stability of Formulation A Containing 100 mg/mL mAb1 AfterStorage at 5° C. in 5 mL Clear Vials t = 0 Storage at 5° C. (months)Parameter 0 1 2 3 6 9 12 Visual Appearance Pass Pass Pass Pass Pass PassPass Turbidity 0.0 0.0 0.0 0.0 0.0 0.0 0.0 (OD 405 nm) pH 6.0 6.1 6.06.1 6.0 6.1 6.1 % Total mAb1 100 106 104 97 100 99 100 Recovered %Native mAb1 96.2 96.3 96.1 96.0 95.9 95.5 95.4 Recovered % Main Peak57.6 57.3 57.9 55.5 56.2 56.4 55.4 mAb1 Recovered % Acidic Species 28.230.2 29.4 31.1 30.7 30.0 32.2 mAb1 Recovered

TABLE 15 Stability of Formulation A Containing 25 mg/mL mAb1 AfterStorage at 5° C. in 5 mL Amber Vials t = 0 Storage at 5° C. (months)Parameter 0 1 2 3 6 9 12 Visual Appearance Pass Pass Pass Pass Pass PassPass Turbidity 0.0 0.0 0.0 0.0 0.0 0.0 0.0 (OD 405 nm) pH 6.0 6.0 6.06.0 6.0 6.0 6.0 % Total mAb1 100 104 104 98 99 97 101 Recovered % NativemAb1 97.1 97.4 96.8 96.7 96.7 96.1 96.3 Recovered % Main Peak 57.8 56.456.3 56.1 55.7 55.9 55.2 mAb1 Recovered % Acidic Species 27.9 30.4 30.631.8 31.0 30.5 32.9 mAb1 Recovered

TABLE 16 Stability of Formulation A Containing 100 mg/mL mAb1 AfterStorage at 5° C. in 5 mL Amber Vials t = 0 Storage at 5° C. (months)Parameter 0 1 2 3 6 9 12 Visual Appearance Pass Pass Pass Pass Pass PassPass Turbidity 0.0 0.0 0.0 0.0 0.0 0.0 0.0 (OD 405 nm) pH 6.0 6.0 6.06.0 6.0 6.0 6.1 % Total mAb1 100 102 104 97 101 100 100 Recovered %Native mAb1 96.2 95.0 96.0 96.0 95.8 95.0 95.3 Recovered % Main Peak57.6 56.9 58.0 55.6 56.3 56.5 55.1 mAb1 Recovered % Acidic Species 28.230.1 29.4 31.4 30.4 30.0 32.3 mAb1 Recovered

TABLE 17 Stability of Formulation A Containing 25 mg/mL mAb1 AfterStorage at 5° C. in 20 mL Amber Vials t = 0 Storage at 5° C. (months)Parameter 0 1 2 3 6 9 12 Visual Appearance Pass Pass Pass Pass Pass PassPass Turbidity 0.0 0.0 0.0 0.0 0.0 0.0 0.0 (OD 405 nm) pH 6.0 6.0 6.06.0 6.0 6.0 6.0 % Total mAb1 100 105 103 97 101 98 101 Recovered %Native mAb1 97.1 96.9 97.0 96.8 97.0 96.2 96.6 Recovered % Main Peak57.8 56.4 57.1 55.9 55.6 56.1 55.2 mAb1 Recovered % Acidic Species 27.920.9 30.2 30.5 30.7 30.0 31.9 mAb1 Recovered

TABLE 18 Stability of Formulation A Containing 100 mg/mL mAb1 AfterStorage at 5° C. in 20 mL Amber Vials t = 0 Storage at 5° C. (months)Parameter 0 1 2 3 6 9 12 Visual Appearance Pass Pass Pass Pass Pass PassPass Turbidity 0.0 0.0 0.0 0.0 0.0 0.0 0.0 (OD 405 nm) pH 6.0 6.0 6.06.0 6.0 6.0 6.0 % Total mAb1 100 105 103 97 99 98 100 Recovered % NativemAb1 96.2 96.2 96.0 96.0 95.8 95.4 95.5 Recovered % Main Peak 57.6 57.956.3 56.5 56.5 56.4 55.3 mAb1 Recovered % Acidic Species 28.2 29.8 30.430.3 30.6 30.0 30.9 mAb1 Recovered

TABLE 19 Stability of Formulation A Containing 100 mg/mL mAb1 AfterStorage at 25° C. and 45° C. in Clear Vials Storage at 45° C. Storage at25° C. t = 0 (days) (months) Parameter 0 7 14 28 1 2 3 6 Visual PassPass Pass Pass Pass Pass Pass Pass Appearance Turbidity 0.0 0.0 0.0 0.00.0 0.0 0.0 0.0 (OD 405 nm) pH 6.0 6.1 6.1 6.1 6.1 6.1 6.1 6.1 % TotalmAb1 100 101 97 105 105 102 97 101 Recovered % Native 95.6 94.8 93.891.2 94.6 93.7 93.1 93.4 mAb1 Recovered % Main Peak 57.6 45.7 34.9 22.353.1 49.0 43.0 37.9 mAb1 Recovered % Acidic 28.2 37.3 49.8 70.8 32.536.8 42.4 53.8 Species mAb1 Recovered

TABLE 20 Stability of Formulation A Containing 100 mg/mL mAb1 AfterStorage at 25° C. and 45° C. in 5 mL Amber Vials Storage at 45° C.Storage at 25° C. t = 0 (days) (months) Parameter 0 7 14 28 1 2 3 6Visual Pass Pass Pass Pass Pass Pass Pass Pass Appearance Turbidity 0.000.00 0.01 0.02 0.00 0.00 0.01 0.02 (OD 405 nm) pH 6.0 6.1 6.1 6.1 6.16.1 6.1 6.0 % Total mAb1 100 101 100 105 106 103 97 100 Recovered %Native mAb1 95.6 94.6 93.6 90.9 94.5 93.7 93.4 92.7 Recovered % MainPeak 57.6 46.1 35.2 21.6 52.7 46.3 42.0 34.2 mAb1 Recovered % AcidicSpecies 28.2 37.3 50.1 70.9 32.8 39.6 43.4 57.3 mAb1 Recovered

TABLE 21 Stability of Formulation A Containing 100 mg/mL mAb1 AfterStorage at 25° C. and 45° C. in 20 mL Amber Vials Storage at 45° C.Storage at 25° C. t = 0 (days) (months) Parameter 0 7 14 28 1 2 3 6Visual Pass Pass Pass Pass Pass Pass Pass Pass Appearance Turbidity 0.000.01 0.00 0.02 0.00 0.01 0.00 0.02 (OD 405 nm) pH 6.0 6.0 6.1 6.1 6.06.0 6.0 6.0 % Total mAb1 100 101 101 105 104 103 96 100 Recovered %Native mAb1 95.6 94.9 93.8 91.5 94.3 94.0 93.6 93.5 Recovered % MainPeak 57.6 45.7 34.2 23.3 52.7 49.3 43.5 36.7 mAb1 Recovered % AcidicSpecies 28.2 36.7 51.1 68.6 32.5 35.8 41.3 55.0 mAb1 Recovered

As shown in this Example, Formulation A containing 25 mg/mL or 100 mg/mLmAb1 exhibited equivalent stability profiles when stored in either clearor amber vials. Moreover, as demonstrated in Example 4 for storage inclear vials, relatively high stability of mAb1 was maintained inFormulation A when stored in either clear or amber vials at 5° C. for upto 12 months.

Example 6 Effect of Arginine, Histidine and Sucrose Concentrations onViscosity and Stability of Formulations Containing 150 mg/mL mAb1

Several formulations were prepared containing 150 mg/mL, 175 mg/mL and200 mg/mL mAb1 and various quantities of histidine, arginine andsucrose. Viscosity and osmolality were measured for each formulation.Additionally, the stability of the 150 mg/mL formulations after 4 weeksof storage at 45° C. was assessed in terms of percent native mAb1remaining (by SE-HPLC) and percent main peak remaining (by CEX-HPLC).The results are summarized in Table 22.

The results presented in Table 16 indicate that increasing the histidineconcentration to 25 mM or 100 mM and adding arginine to the formulation(25 mM or 50 mM) significantly reduced the viscosity of the formulationas compared to formulations containing only 10 mM histidine and noarginine. Furthermore, reducing the sucrose concentration from 10% to 5%with the added histidine and arginine decreased the viscosity of theformulation to an even greater extent.

Based at least in part on the foregoing, the following Formulations(designated “Formulation B” and “Formulation C”) set forth in Table 23were prepared.

TABLE 22 Effect of Arginine, Histidine and Sucrose on Viscosity andStability of mAb1 Formulations % native mAb1 [Histidine] [Arginine][Sucrose] Viscosity Osmolality mAb1 % acidic (mg/mL) (mM) (mM) (%)(cPoise) (mOsm) remaining* species 150 10 0 10 ~15 375 90.8 19.9 150 2525 10 ~11.5 500 91.6 19.5 150 25 25 5 ~8.5 305 91.8 19.8 150 25 25 2.5~8.0 220 91.1 19.9 150 25 25 0 ~8.5 115 90.3 20.6 175 10 0 10 ~27 395175 25 25 10 ~20.5 415 175 25 25 5 ~19 300 175 25 50 5 ~14.5 390 175 1000 5 ~12.5 415 175 100 50 5 ~10 515 200 10 0 10 ~44 410 200 25 25 10 ~35480 200 25 25 5 ~30 300 200 25 25 0 ~28 130 200 100 0 10 ~27 570 200 10050 10 ~21 670 Initial % native mAb1 = 96.5% (SE-HPLC)

TABLE 23 Stabilized mAb1 Formulations “B” and “C” Component FormulationB Formulation C mAb1 25-200 mg/mL 25-200 mg/mL Histidine 25 mM 25 mMPolysorbate 20 0.2% 0.2% Sucrose   5%   5% Arginine 25 mM 50 mM pHadjusted to 6.0

Example 7 Stability of Formulation B Containing 150 mg/mL mAb1 WhenManufactured in a Vial and Syringes

Formulation B (see Table 23) containing 150 mg/mL mAb1 was prepared in a2 mL glass vial and in two different syringes: regular and low tungsten.The preparations were stored at 5, 25 and 45° C. for various amounts oftime. The stability of mAb1 following storage was measured by SE-HPLCand CEX-HPLC. The results are shown in Table 24. (An increase in percentacidic species is consistent with deamidation of the antibody and isthus considered an undesired phenomenon with respect to thepharmaceutical formulations of the present invention).

TABLE 24 Stability of Formulation B Containing 150 mg/mL mAb1 in Vialand Syringe 2 mL Glass Vial Regular Syringe Low Tungsten Syringe %Native % Acidic % Native % Acidic % Native % Acidic Temp Time (SE-HPLC)(CEX-HPLC) (SE-HPLC) (CEX-HPLC) (SE-HPLC) (CEX-HPLC) — Start 96.7 32.296.5 32.3 96.7 31.7 45° C. 14 94.1 52.7 94.3 54.5 94.3 56.1 days 45° C.28 92.7 69.7 92.7 69.7 92.5 70.8 days 45° C. 56 86.7 84.7 87.8 82.6 86.983.5 days 25° C. 1 95.1 31.2 95.7 30.6 95.6 31.1 month 25° C. 2 95.334.6 94.7 37.4 96.0 36.3 month 25° C. 3 94.3 40.6 93.9 43.7 94.1 42.6month  5° C. 1 96.2 30.2 96.5 29.1 96.4 29.3 month  5° C. 2 96.3 29.396.4 29.0 96.4 29.1 month  5° C. 3 95.7 29.4 95.8 29.6 95.8 29.6 month

As shown in this table, Formulation B containing 150 mg/mL mAb1, storedat 5° C. in a glass vial or syringe, remained relatively stable for atleast 3 months.

Example 8 Stability of mAb1 Formulations in Prefilled Syringes

A series of experiments was carried out to assess the stability ofdifferent mAb1 formulations in prefilled syringes. For these experimentsvarious luer and staked needle, regular-tungsten and low-tungstensyringes were used in combination with different types of plungers(coated and uncoated) and tip-caps. The formulations were tested forstability after storage in prefilled syringes at 45° C., 25° C. and 5°C. for various amounts of time (ranging from 14 days to 12 months,depending on the conditions tested).

Six different formulations of mAb1 were tested for stability inprefilled syringes in this Example: (1) Formulation A (see Table 9)containing 100 mg/mL mAb1; (2) Formulation A (see Table 9) containing 25mg/mL mAb1; (3) Formulation B (see Table 23) containing 150 mg/mL mAb1;(4) Formulation B (see Table 23) containing 25 mg/mL mAb1; (5)Formulation C (see Table 23) containing 175 mg/mL of mAb1; and (6)Formulation C (see Table 23) containing 25 mg/mL of mAb1.

Stability was assessed by the following parameters: (a) visual analysis;(b) turbidity (OD405 nm); (c) percent recovery by RP-HPLC; (d) percentnative mAb1 by SE-HPLC; (e) percent main peak mAb1 by CEX-HPLC; and (f)percent acidic species by CEX-HPLC.

The results from a representative experiment assessing the stability ofFormulation A, containing 100 mg/mL mAb1 in two different syringes(Syringe #1 and Syringe #2) are shown in Tables 25 and 26 below.

TABLE 25 Stability of Formulation A containing 100 mg/mL mAb1 in StakedNeedle Prefilled Syringe #1 Syringe #1 Description: Syringe: BD 1 mLlong 29ga × ½″ Physiolis, Low Tungsten Plunger: West FluroTec ® 4023/50Tip Cap: BD 260 Siliconization: Sprayed Visual Turbidity % Native % Main% Acidic Temp Time Analysis (OD_(405 nm)) % Recovery mAb1 Peak Species —Start Pass 0.00 100 96.6 56.9 28.7 45° C. 14 days Pass 0.00 99 95.1 32.750.7 45° C. 28 days Pass 0.01 103 92.6 20.9 66.1 45° C. 56 days Pass0.03 105 88.8 9.9 80.9 25° C. 1 month Pass 0.00 106 95.6 52.4 32.0 25°C. 2 months Pass 0.00 107 95.2 48.0 37.0 25° C. 3 months Pass 0.00 10694.2 44.8 41.8 25° C. 6 months Pass 0.01 101 93.7 34.8 53.9 25° C. 9months Pass 0.03 98 91.4 26.1 64.6 25° C. 12 months Pass 0.03 101 89.921.3 69.4  5° C. 1 month Pass 0.00 110 96.4 56.8 29.9  5° C. 2 monthsPass 0.00 108 96.2 55.7 31.1  5° C. 3 months Pass 0.00 104 96.0 56.330.0  5° C. 6 months Pass 0.00 100 96.5 55.0 31.3  5° C. 9 months Pass0.00 98 96.2 56.7 30.3  5° C. 12 months Pass 0.00 101 95.4 57.3 30.2

The results from another representative experiment assessing thestability of Formulation C, containing 175 mg/mL mAb1 in two differentsyringes (Syringe #1 and Syringe #3) are shown in Tables 27 and 28below.

TABLE 26 Stability of Formulation A containing 100 mg/mL mAb1 in StakedNeedle Prefilled Syringe #2 Syringe #2 Description: Syringe: Schott 1 mLLong SN CF 29ga × ½″ Plunger: West FluroTec ® 4023/50 Tip Cap: Stelmi4800 w/RNS Siliconization: Sprayed Visual Turbidity % Native % Main %Acidic Temp Time Analysis (OD_(405 nm)) % Recovery mAb1 Peak Species —Start Pass 0.00 100 96.3 57.5 28.0 45° C. 14 days Pass 0.00 100 95.233.7 49.6 45° C. 28 days Pass 0.00 103 93.3 22.8 64.7 45° C. 56 daysPass 0.03 107 88.0 9.9 81.1 25° C. 1 month Pass 0.00 108 95.5 52.5 31.825° C. 2 months Pass 0.00 107 95.2 49.2 35.7 25° C. 3 months Fail 0.00106 93.9 43.1 42.0 25° C. 6 months Fail 0.00 102 92.9 34.4 54.0 25° C. 9months Pass 0.02 100 92.3 26.9 63.5 25° C. 12 months Pass 0.03 103 90.020.0 70.2  5° C. 1 month Pass 0.00 111 96.3 56.7 29.9  5° C. 2 monthsPass 0.00 112 95.6 55.9 31.1  5° C. 3 months Pass 0.00 106 96.1 57.229.4  5° C. 6 months Pass 0.00 102 96.0 54.9 31.6  5° C. 9 months Pass0.00 100 95.9 56.7 30.2  5° C. 12 months Pass 0.00 102 95.4 56.0 30.7

TABLE 27 Stability of Formulation C containing 175 mg/mL mAb1 in StakedNeedle Prefilled Syringe #1 Syringe #1 Description: Syringe: BD 1 mLlong 29ga × ½″ Physiolis, Low Tungsten Plunger: West FluroTec ® 4023/50Tip Cap: BD 260 Siliconization: Sprayed Visual Turbidity % Native % Main% Acidic Temp Time Analysis (OD_(405 nm)) % Recovery mAb1 Peak Species —Start Pass 0.00 100 96.7 59.7 32.4 45° C. 7 days Pass 0.01 102 96.1 48.639.5 45° C. 14 days Pass 0.03 97 95.0 36.9 50.0 45° C. 28 days Pass 0.0398 91.9 24.7 66.0 45° C. 56 days Pass 0.05 97 91.9 12.3 83.3 25° C. 1month Pass 0.02 99 95.4 56.9 33.8 25° C. 2 months Pass 0.00 100 95.051.1 39.8  5° C. 1 month Pass 0.00 98 96.1 59.5 32.7  5° C. 2 monthsPass 0.00 101 96.4 56.3 37.1

TABLE 28 Stability of Formulation C containing 175 mg/mL mAb1 in StakedNeedle Prefilled Syringe #3 Syringe #1 Description: Syringe: DaikyoSeiko CZ 1 mL std 30ga × ½″ Plunger: Daikyo D-21-6-1 FluroTec ® CoatedTip Cap: 7028 Siliconization: N/A Visual Turbidity % Native % Main %Acidic Temp Time Analysis (OD_(405 nm)) % Recovery mAb1 Peak Species —Start Pass 0.00 100 96.4 58.2 33.6 45° C. 7 days Pass 0.00 101 95.7 45.440.4 45° C. 14 days Pass 0.01 101 94.8 37.5 48.8 45° C. 28 days Pass0.04  96 94.0 29.4 59.4 45° C. 56 days Pass 0.06  99 85.9  7.8 87.0 25°C. 1 month N/D N/D N/D N/D N/D N/D  5° C. 1 month Pass 0.00 100 96.456.7 34.0  5° C. 2 months Pass 0.00 101 96.2 54.7 34.0

The results from this set of experiments demonstrate that the differentformulations remain relatively stable in prefilled syringes, especiallywhen stored at temperatures of 25° C. and below, for one month orgreater. Moreover, the various formulations of the invention appeared tohave enhanced stability when contained in low tungsten syringescontaining fluorocarbon-coated plungers.

The present invention is not to be limited in scope by the specificembodiments describe herein. Indeed, various modifications of theinvention in addition to those described herein will become apparent tothose skilled in the art from the foregoing description and theaccompanying figures. Such modifications are intended to fall within thescope of the appended claims.

1-31. (canceled)
 32. A stable pharmaceutical formulation comprising: (i)a human antibody that specifically binds to human interleukin-6 receptor(hIL-6R), wherein the antibody is at a concentration of from about 25mg/ml to about 200 mg/ml and comprises a heavy chain variable regionhaving the amino acid sequence of SEQ ID NO:18 and a light chainvariable region having the amino acid sequence of SEQ ID NO:26; (ii)histidine at a concentration of from about 10 mM to about 25 mM; (iii)arginine at a concentration of from about 25 mM to about 50 mM; (iv)sucrose in an amount of from about 5% to about 10% w/v; and (v)polysorbate in an amount of from about 0.1% to about 0.2% w/v, whereinthe formulation has a pH of about 5.8, about 6.0, or about 6.2, and atleast 90% of the native form of the antibody is recovered after 1 monthof storage at 45° C., as determined by size exclusion chromatography.33. The pharmaceutical formulation of claim 32, wherein the histidine isat a concentration of 21 mM.
 34. The pharmaceutical formulation of claim33, wherein the arginine is present at a concentration of 45 mM.
 35. Thepharmaceutical formulation of claim 34, wherein the sucrose is presentin an amount of 5% w/v.
 36. The pharmaceutical formulation of claim 35,wherein the polysorbate is present at a concentration of 0.2% w/v. 37.The pharmaceutical formulation of claim 36 that has a pH of
 6. 38. Thepharmaceutical formulation of claim 32, comprising from 50 mg/mL to 180mg/mL of the human antibody that specifically binds to hIL-6R.
 39. Thepharmaceutical formulation of claim 38, comprising 150 mg/mL of saidhuman antibody that specifically binds to hIL-6R.
 40. The pharmaceuticalformulation of claim 38, comprising 175 mg/mL of said human antibodythat specifically binds to hIL-6R.
 41. The pharmaceutical formulation ofclaim 40, wherein at least 90% of the native form of said antibody isrecovered after nine months of storage at 5° C., as determined by sizeexclusion-high performance liquid chromatography (SE-HPLC).
 42. Thepharmaceutical formulation of claim 41, wherein at least 95% of thenative form of said antibody is recovered after nine months of storageat 5° C., as determined by size exclusion-high performance liquidchromatography (SE-HPLC).
 43. The pharmaceutical formulation of claim42, wherein at least 96% of the native form of said antibody isrecovered after nine months of storage at 5° C., as determined by sizeexclusion-high performance liquid chromatography (SE-HPLC).
 44. Thepharmaceutical formulation of claim 40, wherein the formulation exhibitsa viscosity of less than about 15 cPoise.
 45. The pharmaceuticalformulation of claim 40, wherein the formulation exhibits a viscosity ofless than about 12 cPoise.
 46. The pharmaceutical formulation of claim40, wherein the formulation exhibits a viscosity of less than about 9cPoise.
 47. The pharmaceutical formulation of claim 32, wherein theformulation is contained in a glass vial.
 48. The pharmaceuticalformulation of claim 32, wherein the formulation is contained in asyringe.
 49. The pharmaceutical formulation of claim 32, wherein theformulation is contained in a microinfusor.
 50. The pharmaceuticalformulation of claim 48, wherein said syringe comprises afluorocarbon-coated plunger.
 51. The pharmaceutical formulation of claim48, wherein said syringe is a low tungsten syringe.
 52. A stablepharmaceutical formulation comprising: (i) 25 to 200 mg/mL of a humanantibody that specifically binds to human interleukin-6 receptor(hIL-6R), wherein the antibody comprises a heavy chain variable regionhaving the amino acid sequence of SEQ ID NO:18 and a light chainvariable region having the amino acid sequence of SEQ ID NO:26; (ii)about 21, 22, 23, 24 or 25 mM histidine; (iii) about 3%, 4% or 5% w/vsucrose; (iv) about 0.18%, 0.19% or 0.2% w/v polysorbate 20; and (v)about 45 or 50 mM arginine, wherein the formulation has a pH of about 6,and at least 90% of the native form of the antibody is recovered after 1month of storage at 45° C., as determined by size exclusionchromatography.
 53. A stable pharmaceutical formulation comprising: (i)about 150 mg/mL of a human antibody that specifically binds to humaninterleukin-6 receptor (hIL-6R), wherein said antibody comprises a heavychain and light chain variable region (HCVR/LCVR) amino acid sequencepair of SEQ ID NOs:18/26; (ii) about 21 mM histidine; (iii) about 5%sucrose; (iv) about 0.2% polysorbate 20; and (v) about 45 mM arginine,wherein the formulation has a pH of about 6, and at least 90% of thenative form of the antibody is recovered after 1 month of storage at 45°C., as determined by size exclusion chromatography.
 54. A stablepharmaceutical formulation comprising: (i) about 175 mg/mL of a humanantibody that specifically binds to human interleukin-6 receptor(hIL-6R), wherein said antibody comprises a heavy chain and light chainvariable region (HCVR/LCVR) amino acid sequence pair of SEQ IDNOs:18/26; (ii) about 21 mM histidine; (iii) about 5% sucrose; (iv)about 0.2% polysorbate 20; and (v) about 45 mM arginine, wherein theformulation has a pH of about 6, and at least 90% of the native form ofthe antibody is recovered after 1 month of storage at 45° C., asdetermined by size exclusion chromatography.
 55. The pharmaceuticalformulation of claim 54, wherein the formulation is contained in astacked needle prefilled syringe.
 56. The pharmaceutical formulation ofclaim 54, wherein at least 96% of native form of said antibody isrecovered after two months of storage at 5° C., as determined by sizeexclusion-high performance liquid chromatography.